This proposal addresses broad Challenge Area (06) 'Enabling Technologies'and specific Challenge Topic 06-AA-106, 'Technology Development for Analysis of Alcohol-Related Neural Circuits'. The applicants of this proposal have been conducting a successful interdisciplinary collaboration addressing the very scientific and technological challenges this Topic calls attention to. Clearly, advanced technologies for the analysis of alcohol-related neural circuits and dynamics have been underutilized in spite of the immense amount of data on the alcohol-affected molecular and cellular mechanisms that have been accumulated during the past two decades. Using state-of-the-art electrophysiological and computational techniques in the research of alcohol-impacted neural circuits would provide the opportunity for a better understanding of the circuit- and systemic level mechanisms that are closely related to the behavioral effects of addiction and withdrawal. In fact, this is the primary objective of our proposal. Work described under the two Specific Aims will include development and optimization of novel technologies previously not utilized in the research of alcohol-related neural circuits. Focusing on one the extended amygdala, one of the most important areas in the motivational/reward system of the brain, we propose a series of experiments with sophisticated electrophysiological methods that will shed light on the neural mechanisms of alcohol dependence and withdrawal from a systems neuroscience perspective. In the first Specific Aim we will characterize the integrative and dynamical properties of neurons in the juxtacapsular bed nucleus of stria terminalis (jcBNST, a central element of the extended amygdala circuits) in relation to their intrinsic biophysical properties both in normal and in dependent animals. Next we will characterize how neuromodulatory inputs known to play important roles in alcohol dependence change the dynamical responses of jcBNST neurons. Here, we will use a computer-based electrophysiological technique referred to as dynamic clamp. This technique will allow us to analyze cellular dynamics and circuit interactions with great accuracy and temporal resolution while maintaining the neurons in a high conductance state, smilar to that in the intact brain. The novelty of this approach is that the dynamic clamp will show how they function in an active, temporally complex synaptic environment and how alcohol-related changes in their intrinsic properties impact their output. We will also be able to detect and analyze sophisticated forms of neural computation, such as cellular resonance and pattern selectivity previously not investigated in details. In the second Specific Aim we will study the effects of alcohol at circuit-level interactions, plasticity and modulation by dopamine and corticotropin releasing factor. For these experiments we will build a new electrophysiological system capable of simultaneously recording the spike activity of tens of neurons and delivering spatiotemporally structured stimulus patterns into the neural circuit. This multielectrode system will allow us to observe alcohol-related changes in the synaptic connectivity, signal transduction within the jcBNST and its overall network output to downstrem neuron populations. While in this project we will focus on a specific brain area that is known to be among the most impacted by alcohol and drugs of abuse, the wide applicability of the techniques we utilize will open possibilities for research addressing scientific and health problems related to other functions of the brain. This will strengthen the long-term impact of our research. In addition to the scientific benefits, this project will contribute to the development of the regional and national economy through innovation and job creation. Future economic security will be promoted through the immediate employment of an early stage investigator (the principal investigator) and two postdoctoral scholars. The innovative research program described in this proposal will open further possibilities for collaborations and the involvement of more motivated young scientists. The long term impact will be the generation of new knowledge providing an important contribution to the identification of therapeutical targets for the treatment of neural/behavioral disorders associated with alcohol addiction as well as to the development of new therapeutical targets for relapse. The proposed research will also contribute to possible development of new, innovative technologies applicable to biological systems and scientific challenges beyond the scope of the current project. Using methodology recently developed by CBRE Consulting Inc. in a recent evaluation of the impact of UCSD on the economy, this project is estimated to create 9 new jobs and result in approximately $1 million in total spending in California. Considering the dynamic interaction of the subcontracting institution with the local and national economy, a similar estimation can be made for The Scripps Research Institute. PUBLIC HEALTH RELEVANCE: This project aims to achieve a better understanding of how alcohol addiction and withdrawal affects neuronal communication in the central nervous system at levels of single neurons and neural circuits. Specifically, we will develop and optimize technologies for the study of alcohol-related dysfunctions of neural circuits that will greatly contribute to a better understanding of the pathogenesis of alcohol addiction. Ultimately, this project will contribute to the development of novel therapeutical concepts providing additional socio-economic benefit.